Screening expression libraries with MAbs

Michael Holland M.Holland at dwe.csiro.au
Tue Nov 9 20:00:08 EST 1993


Hi
We have used a monoclonal antibody to an unknown cell surface component to
screen a lambda gt11 cDna library constructed from the tissue which secretes
the protein recognised by the MAb. In the process we obtained a number of
positive clones which went through 3 rounds of screening as positives. After
sequencing and characterisation it became apparent that some clones were
true positives ie they did encode the MAb epitope but others were false ie
they did NOT encode the MAb epitope. How commom is this form of false
positive? Does anyone have similar examples to share? Has anyone seen a
published discussion of this? Why do such false positives survive repeated
screening?
PS The Mab epitope is NOT conformational.
Michael K Holland PhD
CSIRO, Wildlife & Ecology
PO Box 84, Lyneham ACT 2602, Australia
Fax: 61-6-242-9242  Phone:61-6-242-1793
M.Holland at dwe.csiro.au



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